PubMed · 7038661
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M R Keighley. 1982-01-19. Pseudomembranous colitis.. https://pubmed.ncbi.nlm.nih.gov/7038661/
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Human basic fibroblast growth factor (bFGF) was produced from recombinant Escherichia coli by high-cell-density cultivation. In order to develop a purification strategy for large-scale purification, chromatographic sorbents with different anionic functional groups were compared in terms of selectivity for bFGF and stability under cleaning in place (CIP) conditions. Heparin-Sepharose CL-6B, Fractogel EMD-SO3- 650 (S) and SP-Sepharose (high performance) were found suitable for this purpose with decreasing selectivity in that order. Each sorbent was treated eight times under CIP conditions employing both 0.2 and 1.0 M NaOH, in order to study modifications of these sorbents. Heparin-Sepharose displayed more than 50% loss of capacity after the first CIP treatment and decreasing selectivity with each cycle. Both cation exchangers displayed almost constant results regarding selectivity and capacity. The Fractogel EMD-SO3- exhibited only slightly lower selectivity for bFGF than Heparin-Sepharose and the highest capacity of all sorbents tested. Agglomeration of bFGF at low salt concentrations was a serious problem. By direct application of pooled fractions from Fractogel EMD-SO3- onto Heparin-Sepharose a highly pure product was obtained; however, the recovery after Heparin-Sepharose was only 30%.
Lithium, sodium, potassium, and ammonium salts of the industrial polysaccharide gellan were prepared. The salts were freely soluble in water at room temperature (25 degrees C). The opinion had been generally held that heating to 100 degrees C was necessary for gellan to achieve complete solubility in the presence of mono- or multivalent cations. Then, upon cooling, the solutions would form gels. These conclusions were based on the properties imposed upon gellan samples by the presence of contaminating divalent cations. Commercial gellan samples contain calcium and magnesium at levels exceeding 0.9%, sufficient for counterion formation with over one-third of gellan's carboxyl groups. Purification was rapid and included sequential treatments with a cation-exchange (H+) resin, LiOH, NaOH, KOH, or NH4OH, and an anion-exchange (Cl-) resin. About 95% of the divalent cations and nearly 90% of the phosphate that contaminated commercial gellan were removed. The purified monovalent salts of gellan set in the presence of divalent cations and provide well-defined agents for gelling media used for propagation of microbes and plants. In a manner analogous to sodium alginate, solutions of lithium, sodium, potassium, or ammonium gellanate form beads when dropped into solutions of divalent cations. This property was exploited for entrapment of enzymes and cells in beads.
Nitric oxide (NO) generation in a series of 20 burn patients was studied with a novel anion exchange high-performance liquid chromatographic method for the simultaneous determination of nitrite (NO2-) and nitrate (NO3-), the stable metabolic endproducts of NO. The NO values within our survivor group (n = 17) were significantly altered at days 1, 6, and 12 postburn in contrast to controls (n = 23) (p < or = 0.03). NO2- values were significantly depressed in both plasma and urine, whereas NO3- values were significantly elevated in contrast to control values (p < or = 0.03). The ratio of NO2-:NO3- was significantly lower for burn patients versus controls in both plasma and urine (p < 0.01). The NO generation seemed in part to be dependent on the percentage of total body surface area burn, most dramatically elevated in patients with burns of 10 to 40% total body surface area. In subjects who did not survive beyond 36 hours postinjury because of irreversible shock (n = 3), the production of NO was significantly depressed in contrast to survivors and controls (p < 0.0001). However, the NO2-:NO3- ratio (0.001) was relatively unchanged, with reflection of a global depression in NO formation with no change in the individual component release. Burn injury resulted in an increased release/production of NO that in the first postburn week is maximally elevated immediately postinjury. NO release, although decreased at day 6 relative to the day 1 values, remained elevated into the second week postinjury when there was evidence for a further increase in NO production. The enhanced NO3- formation may well result from NO reacting with oxygen-free radicals counteracting superoxide anion-induced destruction of tissue, thereby potentially functioning as a protectant molecule.